新型基因载体纳米儿茶素-Fe^(2+)对人血管平滑肌细胞的转染效率及毒性评价

Transfection efficiency and toxicity of a novel gene carrier nano catechin-Fe^(2+) on human vascular smooth muscle cells

目的探讨纳米儿茶素-Fe^(2+)对人心血管平滑肌细胞的基因转染效率及细胞毒性,为后续对其转染机制及转染条件优化的研究提供依据。方法制备纳米儿茶素-Fe^(2+)的基因导入材料,将质量比为50∶1的儿茶素-Fe^(2+)/pEGFP-C1复合物转染人心血管平滑肌细胞,并设Lipo2000为对照,在荧光显微镜下观察细胞转染情况,采用MTT法检测儿茶素-Fe^(2+)/pEGFP-C1复合物转染对人心血管平滑肌细胞生长的影响,评价细胞毒性。结果纳米儿茶素-Fe^(2+)与pEGFP-C1最佳结合效率为50∶1;纳米儿茶素-Fe^(2+)组与Lipo2000组转染率差异无统计学意义(P>0.05);儿茶素-Fe^(2+)纳米粒子对转染细胞无明显细胞毒性;儿茶素-Fe^(2+)/pEGFP-C1纳米粒子转染后细胞荧光及EGFP荧光强度与Lipo2000组比较,转染效率相当(P>0.05)。结论儿茶素-Fe^(2+)是一种低细胞毒性,高转染率的纳米材料,具备良好的应用前景。

Objective To explore the novel catechin-Fe2＋nanoparticle system,and evaluate its toxicity and transfection efficiency on vascular smooth muscle cells. Methods The novel catechin-Fe2＋nanoparticle system was constructed. The DNA-binding efficiency of catechin-Fe2＋nanoparticle,catechin-Fe2＋and pEGFP-C1 plasmid DNA were determined by use of gel retardation assay. Based on the results of gel retardation assay ,the ratio of 50：1 was considered as the optimal mass ratio. Catechin-Fe2＋/pEGFP-C1 nanoparticles complex were transfected into vascular smooth muscle cells and cells transfected with pEGFP-C1 using lipofectamine2000 was served as a control. Cell transfection was observed under fluorescent microscope,and influence of lentivirus transfection on proliferation of vascular smooth muscle cells was detected by MTT . Results The optimal mass ratio for atechin-Fe2＋ and atechin-Fe2＋ is 50：1. There was no statistical significant difference between the catechin-Fe2＋ transfected and lipofectamine 2000 transfected cells（P＞0.05）. Atechin-Fe2＋ nanoparticles is no significant cell toxicity to transfected cells. Moreover, the cell fluorescence and EGFP fluorescent intensity detected by flow cytometry in the catechin-Fe2＋/pEGFP-C1 nanoparticles transfected cells was as high as lipofectamine 2000 transfected cells（P＞0.05）. Conclusion Catechin-Fe2＋ nanoparticles is a promising system with high transfection efficiency and little toxicity.